JPS631292B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to stable lysozyme or its acid salt containing formulations. More specifically, the present invention relates to a preparation containing lysozyme or its acid salt containing an antioxidant. Lysozyme or its acid salt has anti-inflammatory, antiviral, and antibacterial effects, and preparations containing these are used as preservatives for medicines and foods. The activity of this lysozyme or its acid salt may be impaired depending on the type of excipient used during formulation. For example, common excipients such as polyethylene glycol analogues, polyoxyethylene polymerized nonionic surfactants, unsaturated fatty acids, unsaturated aliphatic alcohols, or unsaturated fatty acid or unsaturated aliphatic alcohol residues. Animal and vegetable oils and fats, waxes, synthetic esters, surfactants, and the like that have the following properties significantly reduce the activity of lysozyme or its acid salts. Therefore, when formulating lysozyme or its acid salt, the excipients to be used are limited, and it is difficult to freely obtain a formulation of desired quality or a desired dosage form. The present inventors investigated the cause of the decrease in activity of lysozyme or its acid salt due to excipients, and attempted to prevent the decrease in activity. As a result, it was confirmed that lysozyme or its acid salts are deactivated by peroxides that are initially contained in excipients or are generated over time, and that deactivation can be prevented or suppressed by adding antioxidants. did. The present invention is a preparation containing lysozyme or its acid salt containing an antioxidant. As the antioxidant, for example, tocopherol, butylated hydroxyanisole (BHA), dibutylated hydroxytoluene (BHT), gallic acid ester, vitamin C higher fatty acid ester, nordihydroguaiaretic acid (NDGA), etc. can be used. The appropriate amount of antioxidants to be added varies depending on the type of antioxidant and the type and amount of excipients, and can be determined by measuring lysozyme activity or peroxide value for each formulation. can. The antioxidant may be formulated with other excipients by a conventional method, but preferably, the antioxidant and peroxide-containing excipient are mixed first, and then lysozyme or its acid salt is mixed. do. EXAMPLES Next, the present invention will be explained in more detail with reference to Examples. The residual rate of lysozyme chloride in the examples measures lysozyme activity [Proceedings of the Society for Experimental Biology and Medicine (Proc.
Soc.Exptl.Biol.Med) Vol. 89, p. 401 (1955)]. Example 1 Ointment 35 parts by weight of polyethylene glycol (4000) and 60 parts by weight of polyethylene glycol (400) at about 75°C
(Peroxide value: 4.62 meq/g).
To this, 0.005, 0.01, 0.05 and 0.1 parts by weight of BHT were added, and the mixture was stirred for 20 minutes while being maintained at about 75°C. The mixture was then cooled to room temperature, and the peroxide value of each base was measured. Each base was heated to 50° C., 5 parts by weight of lysozyme chloride was added, and while uniformly mixed, the mixture was cooled to room temperature and filled into an airtight container to prepare an ointment. After storing this at 45°C for 2 months, the residual titer of lysozyme was measured to determine the residual rate of lysozyme chloride. In addition, as a control, one without BHT was selected. The results are shown in Table 1.

[Table] As shown in the table, as the BHT content increases,
The peroxide value of the base is decreasing, and the residual rate of lysozyme chloride in the ointment is increasing. This is because the cause of the deactivation of lysozyme chloride is the peroxide in the base, and by adding BHT, the peroxide in the base is reduced, which prevents the deactivation of lysozyme chloride. It shows. Example 2 Ointment 15 parts by weight of liquid paraffin, 42 parts by weight of white petrolatum, 8 parts by weight of solid paraffin, 15 parts by weight of glyceryl monooleate, and 15 parts by weight of olive oil at about 75°C.
(Peroxide value: 8.21 meq/g).
Add 0.005 vitamin C monostearate to this,
0.02, 0.05, 0.1 and 0.2 parts by weight were added and stirred for 20 minutes while maintaining the same temperature. The mixture was then cooled to room temperature, and the peroxide value of each base was measured.
Each base was heated to 50° C., 5 parts by weight of lysozyme chloride was added, and while uniformly mixed, the mixture was cooled to room temperature and filled into an airtight container to prepare an ointment. 2 at 45℃
After storage for several months, the residual titer of lysozyme was measured and the residual rate of lysozyme chloride was determined. In addition, as a control, one without the addition of vitamin C monostearate was selected. The results are shown in Table 2.

[Table] As shown in the table, as the amount of vitamin C monostearate increases, the residual rate of lysozyme chloride also increases. Example 3 Ointment Lysozyme chloride 1.0 parts by weight Polyethylene glycol 4000 40.0 BHA 0.01-0.05 Polyethylene glycol 400 Total amount 100.0 An ointment was produced in the same manner as in Example 1. 45℃
The residual rate of lysozyme chloride after storage for one month was determined. As a control, one without added BHA was selected.
The results are shown in Table 3.

[Table] As shown in the table, the residual rate of lysozyme chloride increases with the addition of BHA. Example 4 Ointment Lysozyme chloride 5.0 parts by weight Solid paraffin 8.0 Liquid paraffin 15.0 Purified lanolin 12.0 Glyceryl monooleate 15.0 Olive oil 10.0 Vitamin C monopalmitate 0.15 White petrolatum Total amount 100.0 An ointment was produced in the same manner as in Example 2. After this ointment was stored at 45°C for 2 months, the residual rate of lysozyme chloride was 100.3%. On the other hand, the residual rate of lysozyme chloride was 71.0% in the case where vitamin C monopalmitate was not added. Example 5 Lozenge (composition in one tablet) Lysozyme chloride 5.0 mg Lactose 60.0 White sugar 1000.0 Polyethylene glycol 6000 50.0 Polyvinyl pyrrolidone 8.0 Magnesium stearate 5.0 BHT 0.3 Propyl gallic acid 0.2 Polyethylene glycol 6000 was added to about 80°C. The mixture was warmly melted, BHT and gallic acid propyl ester were added, and the mixture was stirred at the same temperature for about 20 minutes. Next, polyvinyl pyrrolidone was added to this and dissolved with stirring.
The temperature was maintained at approximately 65°C. Lactose, white sugar, and lysozyme chloride were mixed and ground, and the above solution was added thereto and kneaded, followed by granulation and sizing. Next, magnesium stearate was mixed and tableted to make a troche. After storing this lozenge at 45°C for 2 months, the residual rate of lysozyme chloride was 99.5%. On the other hand, in the case where BHT and propyl gallic acid were not added, the residual rate of lysozyme chloride was 72.0%. Example 6 Soft capsule suppositories (composition in one ball) Silicic anhydride 15.0 mg Glyceryl monostearate 25.0 Glyceryl monooleate 100.0 Soy lecithin 50.0 Lysozyme chloride 5.0 Liquid paraffin 920.0 Natural tocopherols (originated from soybeans, containing 88% total tocopherols) 0.2 Vitamin C monopalmitate 0.2 Lysozyme chloride and excipients other than silicic anhydride were mixed and heated to about 70° C. for about 20 minutes to uniformly dissolve. After adding silicic anhydride to this and dispersing it,
After cooling to about 50°C, lysozyme chloride was added, mixed uniformly, and then cooled to room temperature. Next, this liquid was filled into a gelatin soft capsule film by a conventional method to obtain a soft capsule suppository. This soft capsule suppository
The residual rate of lysozyme chloride after storage at 45℃ for 2 months is
It was 97.5%. On the other hand, in the case where natural tocopherol and vitamin C monopalmitate were not added, the residual rate of lysozyme chloride was 78.3%.

Claims (1)

[Scope of Claims] 1. A preparation containing lysozyme or its acid salt and containing an antioxidant. 2. Lysozyme or its acid salt-containing preparation according to claim 1, wherein the antioxidant is one or more selected from tocopherol, butylated hydroxyanisole, dibutylated hydroxytoluene, gallic acid ester, and vitamin C higher fatty acid ester. 3. The preparation containing lysozyme or its acid salt according to claim 2, wherein the gallic acid ester is gallic acid propyl ester. 4 Vitamin C higher fatty acid ester is vitamin C
The lysozyme or its acid salt-containing preparation according to claim 2, which is monostearate or vitamin C monopalmitate.